Variation In Metabolites Research Articles

Comparison analysis of full-spectrum metabolomics revealed on the variation of potential metabolites of unscented, Chloranthus spicatus scented, and Osmanthus fragrans (Thunb.) Lour. scented Congou black teas.

In recent years, scented black tea has attracted much attention due to its pleasant floral aroma and mellow flavor, but little research has been carried out on its flavor metabolic profile. In this study, the flavor metabolic profiles of unscented, Chloranthus spicatus scented, and Osmanthus fragrans (Thunb.) Lour. scented Congou black teas were investigated using full-spectrum metabolomics analysis method, the first time that the flavor profiles of scented black tea were characterized in detail. The results revealed that a total of 3,128 metabolites were detected in the three teas. Based on the criteria of variable importance in the project >1 and fold change ≥2 or ≤ 0.5, 761 non-volatile metabolites and 509 volatile metabolites were filtered as differential metabolites. Many differential non-volatile metabolites belonged to flavonoids, phenolic acids, and terpenoids. Floral, fruity and herbaceous volatile metabolites were significantly up-regulated in Chloranthus spicatus scented Congou black tea while sweet and fruity volatile metabolites were significantly down-regulated in Osmanthus fragrans (Thunb.) Lour. scented Congou black tea. The results contribute to a better understanding of the scenting techniques on the flavor quality of scented black teas and provide some information on the flavor chemistry theory of scented black teas.

Resistance in Soybean Genotypes to Whitefly <i>Bemisia Tabaci</i>

This study demonstrates the defensive responses of eight soybean genotypes based on plant metabolites to the whitefly Bemisia tabaci. At two sampling intervals (30 and 50 days after sowing) secondary metabolites viz., total phenols, o-dihydroxy phenols, flavonols and tannins and primary metabolites viz., total soluble sugars, reducing sugars and total soluble proteins have been estimated from uninfested and infested soybean plants (125 whitefly adults/plant). The genotypes revealed significant variation in secondary metabolites while primary metabolites decreased. The correlation between whitefly and metabolites showed that secondary metabolites were significantly negatively correlated incidence while primary metabolites were significantly positively correlated.

Unveiling the Impact of Different Nitrogen Fertilizer Levels on Rice’s Eating Quality through Metabolite Evaluation

We investigated the variations in metabolites associated with the quality of rice consumption when exposed to varying nitrogen fertilizer levels, as well as the regulatory role of pivotal metabolites within metabolic pathways. This research employed Hongyang 5 as the subject of experimentation, examining the metabolites of Hongyang 5 at three different nitrogen levels using non-targeted metabonomic analysis. The findings indicated that the overall assessment of the eating quality/palatability (CEQ) and amylose contents (AC) of Low nitrogen (D1: 180 kg·ha−1) was notably greater than that of Medium nitrogen (D2: 270 kg·ha−1) and High nitrogen (D3: 315 kg·ha−1). Conversely, the amylopectin (APC), total starch (SC), and protein contents (AP) of D1 were remarkably lower than those observed in D2 and D3. The starch debranching enzyme (DBE) and granule-bound starch synthetase (GBSS) of D1 were remarkably higher than those of D2 and D3. The soluble starch synthase (SSS) of D1 was the lowest. The ADP-glucose pyro-phosphorylase (AGP) and starch branching enzyme (SBE) of D3 were remarkably higher than that of D1 and D2. We identified 76 differential metabolites (DMs) between D1 and D2 (20 up-regulated and 56 down-regulated). A total of 88 DMs were identified between D3 and D1 (42 up-regulated and 46 down-regulated). A total of 57 DMs were identified between D3 and D2. Most of the DMs related to rice-eating quality were involved in the lipid metabolic pathway and amino acid metabolic pathway. The essential metabolites within the metabolic pathway are classified as lipid metabolites and are (13(S)-hydroperoxylinolenic acid, PGB2, 3-phosphocholine, 7-epijasmonic acid, 20-carboxyleukotriene B4 and 11-dehydro-thromboxane B2), amino acid metabolites (4-guanidinobutanoic acid, (3R, 5S)-1-pyrroline-3-hydroxy-5-carboxylic acid, citric acid, (S)-2-Acetolactate, L-glutamine, L-2, 4-aminobutyric acid and putrescine). These key metabolites may be affected by nitrogen fertilizer conditions and play critical regulatory roles in the metabolic pathway, resulting in differences in rice eating quality.

Annotation of genes involved in gamma-aminobutyric multiplication during coix germination by metabolome and transcriptome analysis

ABSTRACT Job’s tears (coix) is a widely used grain with rich and balanced nutritional value. In this study, a total of 71 metabolites, including amino acid metabolism, sugar metabolism and fatty acid metabolism, were significantly increased in the germinating coix through liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Significant increase in gamma-aminobutyric (GABA) content during seed germination was detected in the metabolites detected. Transcript profiles generated through high-throughput RNA-sequencing data and 2431 differentially expressed genes (DEGs) were identified. The functional of DEGs were also significantly clustered in glucose metabolism and fatty acid metabolism. In the annotation of metabolite variation and differentially expressed genes, 23 regulatory genes and 10 significantly enriched metabolites for GABA content proliferation during coix germination were identified by association analysis. These results provided a theoretical basis for understanding the germination mechanism of coix and promoting its production and application.

Regional variation in the antibacterial activity of a wild plant, wild garlic (Allium ursinum L.)

Antibacterial activity is a common and highly studied property of plant secondary metabolites. Despite the extensive literature focusing on identifying novel antibacterial metabolites, little work has been undertaken to examine variation in levels of antibacterial activity in any plant species. Here, we used large-scale sampling of leaves of the antibacterial plant, wild garlic (Allium ursinum L.), assembling a set of tissue extracts from 168 plants, with 504 leaves collected and analysed. We assayed extracts for antibacterial activity against Bacillus subtilis and used LC-MS to carry out a chemometric analysis examining variation in individual metabolites, comparing them with several ecological parameters. We found that allicin was the only metabolite which was positively related to antibacterial activity. Soil temperature was a key determinant of variability in the concentrations of many foliar metabolites, however, neither allicin concentrations nor antibacterial activity was related to any of our measured ecological parameters, other than roadside proximity. We suggest that the synthesis of allicin precursors may be largely independent of growing conditions. This may be to ensure that allicin is synthesised rapidly and in sufficiently high concentrations to effectively prevent herbivory and pest damage. This finding contrasts with flavonoids which were found to vary greatly between plants and across sites. Our findings suggest that key biologically active metabolites are constrained in their concentration range compared to other compounds in the metabolome. This has important implications for the development of wild garlic as a health supplement or animal feed additive.

Assessing the Acute Toxicological Effects of Annona muricata Leaf Ethanol Extract on Rats: Biochemical, Histopathological, and Metabolomics Analyses.

Annona muricata is a common plant used in Africa and South America to manage various types of disease. However, there is insufficient toxicological information or published standard available regarding repeated dose animal toxicity data. As part of the safety assessment, we exposed Sprague Dawley rats to an acute oral toxicity of A. muricata. The intent of the current study was to use advanced proton nuclear magnetic resonance (1H NMR) in serum and urinary metabolomics evaluation techniques to provide the in vivo acute toxicological profile of A. muricata leaf ethanol extract in accordance with the Organization for Economic Co-operation and Development's (OECD) 423 guidelines. A single 2000 mg/kg dose of A. muricata leaf ethanol extract was administered to Sprague Dawley rats over an observational period of 14 days. The toxicity evaluation (physical and behavior observation, body weight, renal function test, liver function test and 1H NMR analysis) showed no abnormal toxicity. Histopathological analysis manifested mild changes, i.e., the treated kidney manifested mild hypercellularity of mesangial cells and mild red blood cell congestion. In addition, there was mild hemorrhage into tissue with scattered inflammatory cells and mild dilated central vein with fibrosis in the liver. However, the changes were very mild and not significant which correlate with other analyses conducted in this study (biochemical test and 1H NMR metabolomic analysis). On the other hand, urinary 1H NMR analysis collected on day 15 revealed high similarity on the metabolite variations for both untreated and treated groups. Importantly, the outcomes suggest that A. muricata leaf ethanol extract can be safely consumed at a dose of 2000 mg/kg and the LD50 must be more than 2000 mg/kg.

Database of space life investigations and information on spaceflight plant biology.

Extensive spaceflight life investigations (SLIs) have revealed observable space effects on plants, particularly their growth, nutrition yield, and secondary metabolite production. Knowledge of these effects not only facilitates space agricultural and biopharmaceutical technology development but also provides unique perspectives to ground-based investigations. SLIs are specialized experimental protocols and notable biological phenomena. These require specialized databases, leading to the development of the NASA Science Data Archive, Erasmus Experiment Archive, and NASA GeneLab. The increasing interests of SLIs across diverse fields demand resources with comprehensive content, convenient search facilities, and friendly information presentation. A new database SpaceLID (Space Life Investigation Database http://bidd.group/spacelid/ ) was developed with detailed menu search tools and categorized contents about the phenomena, protocols, and outcomes of 459 SLIs (including 106 plant investigations) of 92 species, where 236 SLIs and 57 plant investigations are uncovered by the existing databases. The usefulness of SpaceLID as an SLI information source is illustrated by the literature-reported analysis of metabolite, nutrition, and symbiosis variations of spaceflight plants.In conclusion,this study extensively investigated the impact of the space environment on plant biology, utilizing SpaceLID as an information source and examining various plant species, including Arabidopsis thaliana, Brassica rapa L., and Glycyrrhiza uralensis Fisch. The findings provide valuable insights into the effects of space conditions on plant physiology and metabolism.

Chemical Constituents, Antioxidant, and α-Glucosidase Inhibitory Activities of Different Fermented Gynostemma Pentaphyllum Leaves and Untargeted Metabolomic Measurement of the Metabolite Variation.

To assess the effects of microbial fermentation on Gynostemma pentaphyllum leaves (GPL), four probiotics were used to ferment GPL (FGPL) for 7 days. At different stages of fermentation, changes in the active components and biological activities of FGPL were determined. The findings suggest that short-term fermentation with probiotics can enhance both the content and bioactivity of active components in GPL. However, prolonged fermentation may lead to a decline in these aspects. Among them, the best effect was observed with SWFU D16 fermentation for 2 days. This significantly improved the total phenolic and total flavonoid content, antioxidant capacity, and inhibitory ability against α-glucosidase activity with an increase of 28%, 114.82%, 7.42%, and 31.8%, respectively. The high-performance liquid chromatography (HPLC) analysis results also supported this trend. Untargeted metabolomics analysis revealed metabolite changes between GPL and FGPL and the key metabolites associated with these functional activities. These key metabolites are mainly organic acids, flavonoids, carbohydrates, terpenoids, and other substances. KEGG analysis demonstrated that microbial metabolism in diverse environments and carbon metabolism were the most significantly enriched pathways. Among them, 3-(3-hydroxyphenyl) propanoic acid, d-glucose, gallic acid, gluconic acid, l-lactic acid, and l-malic acid were mostly involved in the microbial metabolism of diverse environmental pathways. In contrast, D-glucose, gluconic acid, and l-malic acid were mainly related to the carbon metabolism pathway. This study revealed the positive effect of probiotic fermentation on GPL and its potential metabolism mechanism, which could provide supporting data for further research.

The circadian clock is required for rhythmic lipid transport in Drosophila in interaction with diet and photic condition

Modern lifestyle is often at odds with endogenously driven rhythmicity, which can lead to circadian disruption and metabolic syndrome. One signature for circadian disruption is a reduced or altered metabolite cycling in the circulating tissue reflecting the current metabolic status. Drosophila is a well-established model in chronobiology, but day-time dependent variations of transport metabolites in the fly circulation are poorly characterized. Here, we sampled fly hemolymph throughout the day and analyzed diacylglycerols (DGs), phosphoethanolamines (PEs) and phosphocholines (PCs) using LC-MS. In wild-type flies kept on sugar-only medium under a light-dark cycle, all transport lipid species showed a synchronized bimodal oscillation pattern with maxima at the beginning and end of the light phase which were impaired in period01 clock mutants. In wild-type flies under constant dark conditions, the oscillation became monophasic with a maximum in the middle of the subjective day. In strong support of clock-driven oscillations, levels of the targeted lipids peaked once in the middle of the light phase under time-restricted feeding independent of the time of food intake. When wild-type flies were reared on full standard medium, the rhythmic alterations of hemolymph lipid levels were greatly attenuated. Our data suggest that the circadian clock aligns daily oscillations of DGs, PEs, and PCs in the hemolymph to the anabolic siesta phase, with a strong influence of light on phase and modality.

Influences of chemotype and parental genotype on metabolic fingerprints of tansy plants uncovered by predictive metabolomics

Intraspecific plant chemodiversity shapes plant-environment interactions. Within species, chemotypes can be defined according to variation in dominant specialised metabolites belonging to certain classes. Different ecological functions could be assigned to these distinct chemotypes. However, the roles of other metabolic variation and the parental origin (or genotype) of the chemotypes remain poorly explored. Here, we first compared the capacity of terpenoid profiles and metabolic fingerprints to distinguish five chemotypes of common tansy (Tanacetum vulgare) and depict metabolic differences. Metabolic fingerprints captured higher variation in metabolites while preserving the ability to define chemotypes. These differences might influence plant performance and interactions with the environment. Next, to characterise the influence of the maternal origin on chemodiversity, we performed variation partitioning and generalised linear modelling. Our findings revealed that maternal origin was a higher source of chemical variation than chemotype. Predictive metabolomics unveiled 184 markers predicting maternal origin with 89% accuracy. These markers included, among others, phenolics, whose functions in plant-environment interactions are well established. Hence, these findings place parental genotype at the forefront of intraspecific chemodiversity. We recommend considering this factor when comparing the ecology of various chemotypes. Additionally, the combined inclusion of inherited variation in main terpenoids and other metabolites in computational models may help connect chemodiversity and evolutionary principles.

Ancestry-driven metabolite variation provides insights into disease states in admixed populations

BackgroundMetabolic pathways are related to physiological functions and disease states and are influenced by genetic variation and environmental factors. Hispanics/Latino individuals have ancestry-derived genomic regions (local ancestry) from their recent admixture that have been less characterized for associations with metabolite abundance and disease risk.MethodsWe performed admixture mapping of 640 circulating metabolites in 3887 Hispanic/Latino individuals from the Hispanic Community Health Study/Study of Latinos (HCHS/SOL). Metabolites were quantified in fasting serum through non-targeted mass spectrometry (MS) analysis using ultra-performance liquid chromatography-MS/MS. Replication was performed in 1856 nonoverlapping HCHS/SOL participants with metabolomic data.ResultsBy leveraging local ancestry, this study identified significant ancestry-enriched associations for 78 circulating metabolites at 484 independent regions, including 116 novel metabolite-genomic region associations that replicated in an independent sample. Among the main findings, we identified Native American enriched genomic regions at chromosomes 11 and 15, mapping to FADS1/FADS2 and LIPC, respectively, associated with reduced long-chain polyunsaturated fatty acid metabolites implicated in metabolic and inflammatory pathways. An African-derived genomic region at chromosome 2 was associated with N-acetylated amino acid metabolites. This region, mapped to ALMS1, is associated with chronic kidney disease, a disease that disproportionately burdens individuals of African descent.ConclusionsOur findings provide important insights into differences in metabolite quantities related to ancestry in admixed populations including metabolites related to regulation of lipid polyunsaturated fatty acids and N-acetylated amino acids, which may have implications for common diseases in populations.

The dynamic variations of flavonoid metabolites in flower buds for Zingiber mioga at different developmental stages

Zingiber mioga Roscoe, a perennial herb, is favoured by consumers for its unique flavor and purplish-red appearance. Studies have shown flavonoids, such as anthocyanins, procyanidins, flavonols and flavones, have a great influence on its appearance quality, flavor formation and physiological function. The production and accumulation of those metabolites are particularly closely related to the appearance formation of Z. mioga flower buds. Therefore, the changes in the relative content of anthocyanins, procyanidins and other flavonoids in flower buds of Z. mioga during the whole developmental stage were studied. More importantly, we depicted the association landscape between anthocyanins and procyanidins as well as other flavonoids such as flavones, flavonols and flavanols, which were natural plant compounds that affected the appearance quality and nutritional value of Z. mioga. The results showed that the total anthocyanins and most anthocyanins and procyanidins, as well as the other flavonoids, all reached the highest value at the maturity stage. Among these secondary metabolites, the quercetin-3-O-(2''-O-rhamnosyl) galactoside, quercetin-4′-O-glucuronide, quercetin-3-O-neohesperidoside and sexangularetin-3-O-glucoside-7-O-rhamnoside in flavonoids, the delphinidin-3-O-glucoside, petunidin-3-O-rutinoside, delphinidin-3-O-(6''-O-p-coumaroyl) glucoside and petunidin-3-O-(6''-O-p-coumaroyl) glucoside in anthocyanins, and procyanidin B2 in procyanidins played a major role in these changes in the enhancement of appearance quality and physiological function during its growth. Furthermore, anthocyanins might synergize with procyanidins and other flavonoids to jointly affect the color formation of flower buds in Z. mioga at the development and maturity stages.

Biological Characterization and Metabolic Variations among Cell-Free Supernatants Produced by Selected Plant-Based Lactic Acid Bacteria.

The aim of this research was to assess the antibacterial and antioxidant properties as well as the variation in metabolites of the cell-free supernatant (CFS) produced by lactic acid bacteria (LAB) from local plants: Lactiplantibacillus plantarum ngue16, L. plantarum ng10, Enterococcus durans w3, and Levilactobacillus brevis w6. The tested strains exhibited inhibitory effects against pathogens, including Bacillus cereus, B. subtilis, Cronobacter sakazakii, Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus using the agar spot assay and well diffusion method. The CFS from all four strains displayed antibacterial activity against these pathogens with minimum inhibitory concentration (MIC) values ranging from 3.12 to 12.5 mg/mL and minimal bactericidal concentration (MBC) values ranging from 6.25 to 25.0 mg/mL. Moreover, the CFS demonstrated resilience within specific pH (3-8) and temperature (60-100 °C) ranges and lost its activity when treated with enzymes, such as Proteinase K and pepsin. Furthermore, the CFS exhibited antioxidant properties as evidenced by their ability to inhibit the formation of two radicals (1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) compared to the negative control, De Man, Rogosa, and Sharpe (MRS) broth. The use of proton-based nuclear magnetic resonance (1H-NMR) spectroscopy revealed the presence and quantification of 48 metabolites in both the CFS and MRS broths. Principal Component Analysis (PCA) effectively differentiated between CFS and MRS broth by identifying the specific metabolites responsible for the observed differences. The partial least squares (PLS) model demonstrated a significant correlation between the metabolites in the LAB supernatant and the tested antibacterial and antioxidant activities. Notably, anserine, GABA, acetic acid, lactic acid, uracil, uridine, propylene glycol, isopropanol, serine, histidine, and indol-3-lactate were identified as the compounds contributing the most to the highest antibacterial and antioxidant activities in the supernatant. These findings suggest that the LAB strains investigated have the potential to be utilized in the production of functional foods and the development of pharmaceutical products.

Camel milk peptides alleviate hyperglycemia by regulating gut microbiota and metabolites in type 2 diabetic mice

This study aimed to investigate the hypoglycemic effect of Camel milk peptides (CMPs) on Type 2 diabetes mellitus (T2DM) mice and reveal its related mechanism from the aspect of gut microbiota and metabolites. The administering CMPs significantly alleviated the weight loss, polydipsia and polyphagia, reduced fasting blood glucose (FBG), improved insulin resistance and sensitivity, and restored the level of serum hormones, lipopolysaccharide (LPS), lipid metabolic and tissue damage. Furthermore, CMPs intervention remarkably reversed gut microbiota dysbiosis in T2DM mice by reducing the relative abundance of Proteobacteria, Allobaculum, Clostridium, Shigella and the Firmicutes/Bacteroidetes ratio, while increasing the relative abundance of Bacteroidetes and Blautia. Metabolomic analysis identified 84 different metabolites between T2DM and CMPs-treated groups, participating in three pathways of Pantothenate and CoA biosynthesis, Phenylalanine metabolism and Linoleic acid metabolism. Ureidopropionic acid, pantothenic acid, hippuric acid, hydrocinnamic acid and linoleic acid were identified as key acidic metabolites closely related to hypoglycemic effect. Correlation analysis indicated that CMPs might have a hypoglycemic effect through their impact on gut microbiota, leading to variations in short-chain fatty acids (SCFAs), acidic metabolites and metabolic pathways. These findings suggest that CMPs could be a beneficial nutritional supplement for intervention T2DM.

Plant species richness regulated by geographical variation down-regulates triterpenoid compounds production and antioxidant activities in white birch bark

White birch (Betula platyphylla Suk.) is a pioneer tree species widely distributed in the high northern hemisphere, and its bioactive compounds have ecological and traditional medicinal roles. How ecological factors impact triterpenoid production is not well understood yet. This study examined the effects of geo-climate, soil properties and plant community properties on triterpenoid compounds synthesis and antioxidant activities of white birch bark at eight locations (13° latitude and 6° longitude ranges) in Northeast China. The results showed that the contents of total triterpenoids, betulin and betulinic acid in white birch bark were high at higher latitudes, and low at lower latitudes. The antioxidant activity of the extracts analyzed by DPPH (1, 1-diphenyl-2-picrylhydrazyl), ABTS (2, 2-azinobis-3-ethylbenzothiazoline-6-sulphonic acid), and FRAP (ferric reducing antioxidant power assay) methods varied across different locations and positively correlated with triterpenoid compound contents. Redundancy analysis indicated that more than 40% of the secondary metabolite variations could be explained by plant species richness in the community, largely regulated by geographical variation. Our findings highlighted that higher species richness reduced triterpenoid accumulation and antioxidant potential. At the same time, the harsh environments in cold and dry geo-climate increased triterpenoid production of white birch. This study favors the efficient use of white birch for triterpenoid production and implies that the stress of harsh environment will promote the synthesis of the secondary metabolite synthesis of dominant trees.

Immunometabolic coevolution defines unique microenvironmental niches in ccRCC

Tumor cell phenotypes and anti-tumor immune responses are shaped by local metabolite availability, but intratumoral metabolite heterogeneity (IMH) and its phenotypic consequences remain poorly understood. To study IMH, we profiled tumor/normal regions from clear cell renal cell carcinoma (ccRCC) patients. A common pattern of IMH transcended all patients, characterized by correlated fluctuations in the abundance of metabolites and processes associated with ferroptosis. Analysis of intratumoral metabolite-RNA covariation revealed that the immune composition of the microenvironment, especially the abundance of myeloid cells, drove intratumoral metabolite variation. Motivated by the strength of RNA-metabolite covariation and the clinical significance of RNA biomarkers in ccRCC, we inferred metabolomic profiles from the RNA sequencing data of ccRCC patients enrolled in 7 clinical trials, and we ultimately identifyied metabolite biomarkers associated with response to anti-angiogenic agents. Local metabolic phenotypes, therefore, emerge in tandem with the immune microenvironment, influence ongoing tumor evolution, and are associated with therapeutic sensitivity.

Evaluation of genetic variation of morphological and clubroot-resistance traits of radish and metabonomic analysis of clubroot-resistant cultivar

Global commercial production of radish (Raphanus sativus) is highly threatened by Plamodiophora brassicae. Germplasm evaluation and the deep analysis of variants with clubroot-resistant trait are needed to produce quality cultivars. The genetic diversity was analyzed for 97 radish germplasms based on 20 agronomic traits and the response to clubroot. Additionally, differentially regulated metabolites (DRMs) between healthy and P. brassicae-infected resistant cultivars were measured. The genetic diversity index values of quality traits varied from 0.396 to 1.350, and that of quantitative traits varied from 1.65–2.15. The variation of radish quantitative traits ranged from 19.18% to 54.44%. The principal component analysis indicated 8 comprehensive indicators, allowing classification of germplasms into 4 classes, with 71 in class I, 20 in class II, 4 in class III, and 2 in class IV. The results of classification of germplasms by the response to clubroot infection only or by comprehensive agronomic traits including response of plants to clubroot disease infection were similar. Among all traits assessed in the present study, the clubroot severity and the depth of in-soil root were closely linked. Metabolomic analysis revealed that the metabolites changes of clubroot resistant cultivar “DHBC” pre- and post- P. brassicae infection was relatively stable. P. brassicae infection resulted in limited DRMs, significant down-regulation was identified on ginsenoside, dihydro-2(3H)-thiophenone, proline, 1-pyrroline, while apiosylglucosyl 4-hydroxybenzoate, chrycolide, glucomalcomiin, and phosphocholine were significantly up-regulated. We revealed more extensive genetic variation in quantitative traits than in quality indexes in radish and we summarized that the response of radish to clubroot disease may be a dominate indicator in evaluation of radish varieties. Finally, untargeted metabolomic study confirmed the clubroot resistance of “DHBC” and revealed a sophisticated metabolites (including some precious pharmaceutical ingredients) variation in response to clubroot infection, offers new directions in the studies of clubroot resistance mechanism of radish.

QT–GWAS: A novel method for unveiling biosynthetic loci affecting qualitative metabolic traits

AbstractAlthough the plant kingdom provides an enormous diversity of metabolites with potentially beneficial applications for humankind, a large fraction of these metabolites and their biosynthetic pathways remain unknown. Resolving metabolite structures and their biosynthetic pathways is key to gaining biological understanding and to allow metabolic engineering. In order to retrieve novel biosynthetic genes involved in specialized metabolism, we developed a novel untargeted method designated as qualitative trait GWAS (QT–GWAS) that subjects qualitative metabolic traits to a genome-wide association study, while the conventional metabolite GWAS (mGWAS) mainly considers the quantitative variation of metabolites. As a proof of the validity of QT-GWAS, 23 and 15 of the retrieved associations identified in Arabidopsis thaliana by QT-GWAS and mGWAS, respectively, were supported by previous research. Furthermore, seven gene-metabolite associations retrieved by QT–GWAS were confirmed in this study through reverse genetics combined with metabolomics and/or in vitro enzyme assays. As such, we established that CYTOCHROME P450 706A5 (CYP706A5) is involved in the biosynthesis of chroman derivatives, UDP-GLYCOSYLTRANSFERASE 76C3 (UGT76C3) is able to hexosylate guanine in vitro and in planta, and SULFOTRANSFERASE 202B1 (SULT202B1) catalyzes the sulfation of neolignans in vitro. Collectively, our study demonstrates that the untargeted QT–GWAS method can retrieve valid gene–metabolite associations at the level of enzyme-encoding genes, even new associations that cannot be found by the conventional mGWAS, providing a new approach for dissecting qualitative metabolic traits.

137-OR: Metabolite Ratios and Development of Type 1 Diabetes

Ratios between metabolites more directly reflect flux through chemical reactions than individual metabolites. Variation in metabolites and their ratios are determined, in part, by genetic factors. Thus, genetic loci associated with variation in metabolite ratios may help disentangle genetic from non-genetic processes captured by the metabolome. We integrated metabolite ratios with genetics to predict changes preceding type 1 diabetes (T1D) in The Environmental Determinants of Diabetes in the Young (TEDDY) study. Quantitative trait loci (QTLs) were identified for 132 metabolite ratios previously associated with progression from islet autoimmunity (IA) to T1D. Ratios were calculated from untargeted metabolomics data available at the time of seroconversion to IA for 338 cases and 921 matched controls in TEDDY. Genotyping was performed using a genome-wide array with custom content with imputation to the TOPMed reference panel. Ratio-SNP associations were estimated using Matrix eQTL software, adjusting for case-control status and genetic relatedness. Genome-wide significant association (P < 4.9×10-9) was observed at 202 independent loci, involving 49 metabolite ratios. Twenty-six loci (12.9%) involved ratios between polyunsaturated fatty-acids (PUFAs) and fatty acid esters of hydroxy fatty acids (FAHFAs), a novel class of bioactive lipids with antidiabetic and anti-inflammatory effects. Variation in the metabolite ratio of eicosadienoic acid to FAHFA (18:1/O-15:1) was significantly determined by a site on chromosome 3 (ITPR1); ITPR1 encodes a receptor integral to calcium signaling and glucose-induced insulin secretion. Other genetic loci contributing to variation of PUFA:FAHFA ratios reside in genes controlling inflammatory and innate immune response (e.g., TSPO, TTLL12, NCAM1) and apoptosis (PPM1L). These novel metabolite ratio QTLs, some with known association with T1D risk, expand potential biomarkers of T1D risk, with PUFA:FAHFA ratios representing candidate FAHFA biosynthesis genes. Disclosure R.K. Johnson: None. A. Romero: None. E. Willems: Employee; Amgen Inc. S.S. Rich: None. M. Rewers: Research Support; Provention Bio, Juvenile Diabetes Research Foundation (JDRF), Hemsley Charitable Trust, Dexcom, Inc., Janssen Research & Development, LLC. O. Fiehn: None. J.M. Norris: None. Funding National Institute of Diabetes and Digestive and Kidney Diseases (R03DK127472); National Institute of Allergy and Infectious Diseases; National Institute of Child Health and Human Development; National Institute of Environmental Health Sciences; JDRF; Centers for Disease Control and Prevention

188-OR: Circulating Metabolites and Type 2 Diabetes (T2D) Risk—An Integrated Metabolomics and Genetics Study of ~23,000 Adults

Metabolome captures complex states of metabolic homeostasis affected by genetic and environmental factors. A systematic understanding of metabolites associated with T2D risk may offer potentials for personalized risk monitoring and prevention. Here, we integrated data for 467 harmonized circulating metabolites, genetics, and diet/lifestyle factors in up to 23,616 racially diverse individuals from 10 cohorts with up to 26-y of follow-up. Through multivariable-adjusted metabolome-wide association analyses, we identified 235 metabolites associated with incident T2D (4,000 incident cases; FDR<0.05). Genetic determinants of these metabolites were mapped to genes implicated in pathways of nutrient/energy metabolism and glucose/lipid dysregulation (pathway enrichment FDR<0.05) and highly expressed in T2D-relevent tissues e.g., liver, adipose tissues, pancreas, and muscles (tissue-specific expression enrichment FDR<0.05). Mendelian randomization analyses further corroborated potential causal associations between 93 of the 235 metabolites and T2D. These included previously known (e.g., valine and isoleucine) and novel associations (e.g., α-ketoisocaproate in leucine metabolism, N-acetylaspartate in glutamate metabolism, and odd-chain fatty acid heptadecanoate). Furthermore, we analyzed metabolome-wide associations with 18 mutually adjusted modifiable risk factors. Several factors e.g., physical activity and whole grain (WG) intake, explained higher proportions of inter-individual variation of T2D-associated vs. other metabolites. We identified specific metabolites as potential mediators linking each risk factor to T2D (e.g., 4 metabolites mediated 22% of the WG-T2D association). In conclusion, we found known and novel metabolites associated with T2D risk, indicating potential biological pathways through which genetic and lifestyle factors contribute to T2D. Our findings may inform future personalized T2D prevention. Disclosure J.Li: None. J.Dupuis: None. E.Selvin: None. S.Bhupathiraju: None. J.A.Brody: None. Y.Liu: None. A.Eliassen: None. J.E.Manson: None. C.B.Clish: None. R.N.Lemaitre: None. K.L.Tucker: None. J.Hu: None. J.I.Rotter: None. M.A.Martinez-gonzalez: None. K.M.Rexrode: None. J.B.Meigs: Consultant; Quest Diagnostics. E.Boerwinkle: None. R.Kaplan: None. F.Hu: None. B.Yu: None. Q.Qi: None. N.The nhlbi topmed metabolomics and proteomics worki: n/a. L.Liang: None. M.Guasch: None. J.Merino: None. M.Ruiz-canela: None. K.Luo: None. C.Rebholz: None. B.Porneala: None. Funding National Institutes of Health (R00DK122128 to J.L.)